Influence of methanol/sorbitol co-feeding rate on pAOX1 induction in a Pichia pastoris Mut+ strain in bioreactor with limited oxygen transfer rate

Journal of Industrial Microbiology & Biotechnology
F CarlyPatrick Fickers

Abstract

High Pichia pastoris biomass density could be obtained using high co-feeding rate of methanol and sorbitol in a fed-batch or continuous culture, while further higher feeding rate finally leads to oxygen limitation in bioreactor. In the literature, there is lack of report about AOX1 promoter regulation with regard to dissolved oxygen level (DO). Therefore, in this work, chemostat cultures were performed to investigate the cell growth, metabolism and regulation of the AOX1 promoter (pAOX1) regarding co-feeding rate of optimized methanol/sorbitol mixture (methanol fraction 0.60 C-mol/C-mol) using a P. pastoris Mut+/pAOX1-lacZ strain. The oxygen transfer rates (OTR) in bioreactor were kept in the range of typical values of large bioreactor, i.e., 4-8 g/(L h) if DO equals 30 % saturation or 5-10 g/(L h) if DO nears zero. For DO >0, an increase of the carbon fed led to an increase of pAOX1 induction. By contrast, when dissolved oxygen was completely depleted, methanol accumulated, causing a 30 % decrease of pAOX1 induction. However, this decrease is more likely to be lined to methanol accumulation than to low level of dissolved oxygen (<4 % DO). Methanol/sorbitol co-feeding allowed cells to adapt to oxygen transient limitations that ...Continue Reading

References

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Apr 11, 2003·Journal of Industrial Microbiology & Biotechnology·Wenhui ZhangMichael M Meagher
Apr 28, 2004·Biotechnology and Bioengineering·A E CunhaM J T Carrondo
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May 26, 2009·Nature Biotechnology·Kristof De SchutterNico Callewaert

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Citations

Jan 3, 2019·Critical Reviews in Biotechnology·Wan-Cang LiuPing Zhu
Jun 3, 2018·Brazilian Journal of Microbiology : [publication of the Brazilian Society for Microbiology]·Andrea B ZepedaJorge G Farías
Feb 27, 2017·Brazilian Journal of Microbiology : [publication of the Brazilian Society for Microbiology]·Cesar Andres Diaz AriasAdalberto Pessoa Junior

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